- Email: [email protected]
Autoantibody to carbonic anhydrase in autoimmune disease sera —Its possible clinical and pathogenical significance
386
AN EXONIC POINT MUTATION WAS ELUCIDATED IN NOVEL ANGIOKERATONA CORPORIS DIFFUSUM. TAMOTSU KANZAKIl. ANN M. WANG', AND ROBERT J. DESNICK'. Dept. of Dermatol. Nagoya City Univ. Nagoya , Div. 9f Med. Mol. Genet. Mount Sinai Sch. of Med. New York, N.Y.
POLYMORPHONUCLEAR LEUKOCYTE ACTIVITY AND IhJMUNOPLUORESCENCESTUDIES IN SWEETS SYNDROME AND BEHDTS DISEASE. INOUE CHIYUKI, WATANABE RIB, KAWA YOUKO, AND iwzOGuCHrMASAK0. Department of Dermatology, Teikyo UniversitySchool of Medicine, Tokvo. <-. Janan. Behget’s disease(BD) and Sweet’s syndrome(SS) have very similar clinical and laboratory findings. To find out further immunological conditions of the two diseases, we studied polymorphonuclear leukocyte(PMN) activity by polarization assay and direct immunofluorescence testing of their lesions. N-formyl-meddonyl-leucyl-phenylslanine(PMLP)-mduc&polarization and random polarization wm significantly higher in the SS patients with an active stage than in controls. We also examined SS patients with an inactive stage and polarization responses correlated with the activity of SS. Those results were almost the same as those of BD. Immunohistcchemically, deposits of C3 were demonstrated in all 8 cases of BD, though the degree was variable. However, some results in SS were positive and some were negative. ~I
Novel angiokeratomacorporis diffusum, a lysosomal strage disease, was reported (Lancet, 1989) and the abnormal matabolites in urine were found to be amino acid 0-glycosides (J. Biol.Chem. 1990). This disease was elucidated to be caused by deficienta-N-acatylgalactosainidase. This enzyme is encoded by a gene mapped to 22q13 -,22qter. In order to determine the mCleC"lar nature of this enzyme, efforts were made to isolate the full-length cDNA and genomic sequences of the ellzyme. This enzyme was purified, oligonucleotideprobes were synthesized and a full-length cDNA was isolated. The 2156 bp cDNA encoded 411 amino acids. This enzyme was found to have nine exons and a single C .+ T transition was identified in the eighth won. The C--fT transition leaded to Argfhp at the 329th amino acid of the enzyme. This point mutation resulted in enzymaticallynon-active protein. Thus, this new disease ~8s elucidated to be caused by an exonic point mutation.
ULTRASTRUCTURALAND BIOCHEMICAL STUDIES OF A PATIENT WITH MENTAL RETARDATION AND MULTIPLE ANGIOMA. T.KANZAKIl, Y.KUNOl. Y.HIRASAYPSH12,AND N.G.FASER'. Deot. of Dermatol.. Nanova and Deot. of _- Citv - Univ.._Nanova .ii , -Bibch.rn Univ., of Shizuoka, Shizucka and Raigmore Hosp., scotw?. A Scot
female, 26 y/o, with mental retardation had multiple small angioma on her trunk which was recognized at 8 y/o (J.Med.Genet.). It was capillary hemangioma histologically. Electron microscopic studies disclosed many clear membrane-limitedcytoplasmic vacuoles in endathelial cells, pericytes and dermal fibroblasts. These clear vac"oles have been reported to be observed in various lysosomal storage diseases, such as mucopolysaccharidoses.glyccproteinosesor glycolipidoses. Enzymatic activities of lysosomal enzymes in her serum were within normal limits. Those were A-,Pgalactosidase,slalidase,j-L-fucosidase, d-N-acetylgalactosaminidase,d-rrannosidase,P-glucuronidase,6 -glucosidase and others. Biochemical snalybea with thin layer chromatographies showed no abnormality in glycopeptidesbut disclosed abnormal ganliosides exretion in her urine. These results suggest that this patient appears to have novel gangliosidosiswith mental retardation and multiple capillary hemangioma.
The possible role of fat cellsin the pathogenesis cl cutaneous leSiCnSOf Behcet'ssyndrome: Animmunoelectron microscopicstudy
DongsikBang,MD andSungnackLee.MD Departmentof Dermatology,Yonsei University Seoul. Korea
AUTOANTIBODY TO CARBONIC ANHYDRASE IN AUTOIMMUNE DISEASE SERA -ITS POSSIBLE CLINICAL AND PATHOGENICAL SIGNIFICANCEYASUNORI INAGAKI, YOKO YOSHIDA AND HIROAKI UEKI Department of Dermatology, Kawasaki Medical School, Kurashlki Carbonic anhydrase (CA) is an extremely basic and important enzyme for a regulation of acid-base status in viva with a wide phyletic distribution. Several different classes of this enzyme are known, and, at least, four enzymi’c forms were designated in humans. However, their amino acid sequences show a great deal of homology. CA are widely distributed in mamJKIli.3” tissues such as erythrocytes, gastric mucosa, renal cells, epidermal cells and so on. We found autoantibody reactive with CA, in sera from patients with several autoimmune disease, using the immunoblotting and the ELISA method. The incidence of the antibody “as not high among the patients. Thus we can conclude that occurrance of the autoantibody reactive with such a” important and fundamental enzyme is a significant phenomenon in autoimmune disorders.
PURIFICATION ANTIGEN YOSHINAO
Collegeof Medicine,
AND
NURO,
Department
attention was paidtothe appearance
of lymphocyte-macrophage-fat-cell adhesiOn and fat-cell lysisin the lobulesOf the subcutaneous fat of the erythema nodosum-like lesions of Behcet's syndrome. In this study, biopsy specimens from 10 patientswith typicalBehcet's syndrome were examined by immunoelectrcn microscopy with PaI'fiCUlar attentiontothechange of fat
cells
OF A HUNAN
DNA BINDING
NASARU
CENTRONERE
PROTEIN
OHASHI.
of Dermatology,
Nagoya
Univ.
School
of Medicine,
Nagoya Anticentromere Investlgatc
We have recentlyreported that special
CHARACTERIZATION
AS AN ALPHOID
et al.
reported
antigens, Hela
the
CENP-B,
satellite)(J. from
antibody
has provided
the biochemistry
Call
nuclear
interaction
and
of one
an alphoid
Biol.109:1963). extract
a probe
of the centramere. DNA We
to Nasumoto
of centromere
(a human
centromeric
purified
by its alpboid
DNA
CENP-Ei
binding
activity. Q-sepharose and alphoid DNA column were used, and fractions were assayed by DNA lmmunoprscipitation and immunoblottlng in each step of the purification. Specific activity of alphoid DNA blndinn was Increased over ten thousand fold. The resultsof DNaseI footprinting and south-western blot suggested that CENP-B itself has the activity to bind to a specific sequence of an alphoid DNA. We conclude that CENP-B is a specific DNA binding protein.
AN EXONIC POINT MUTATION WAS ELUCIDATED IN NOVEL ANGIOKERATONA CORPORIS DIFFUSUM. TAMOTSU KANZAKIl. ANN M. WANG', AND ROBERT J. DESNICK'. Dept. of Dermatol. Nagoya City Univ. Nagoya , Div. 9f Med. Mol. Genet. Mount Sinai Sch. of Med. New York, N.Y.
POLYMORPHONUCLEAR LEUKOCYTE ACTIVITY AND IhJMUNOPLUORESCENCESTUDIES IN SWEETS SYNDROME AND BEHDTS DISEASE. INOUE CHIYUKI, WATANABE RIB, KAWA YOUKO, AND iwzOGuCHrMASAK0. Department of Dermatology, Teikyo UniversitySchool of Medicine, Tokvo. <-. Janan. Behget’s disease(BD) and Sweet’s syndrome(SS) have very similar clinical and laboratory findings. To find out further immunological conditions of the two diseases, we studied polymorphonuclear leukocyte(PMN) activity by polarization assay and direct immunofluorescence testing of their lesions. N-formyl-meddonyl-leucyl-phenylslanine(PMLP)-mduc&polarization and random polarization wm significantly higher in the SS patients with an active stage than in controls. We also examined SS patients with an inactive stage and polarization responses correlated with the activity of SS. Those results were almost the same as those of BD. Immunohistcchemically, deposits of C3 were demonstrated in all 8 cases of BD, though the degree was variable. However, some results in SS were positive and some were negative. ~I
Novel angiokeratomacorporis diffusum, a lysosomal strage disease, was reported (Lancet, 1989) and the abnormal matabolites in urine were found to be amino acid 0-glycosides (J. Biol.Chem. 1990). This disease was elucidated to be caused by deficienta-N-acatylgalactosainidase. This enzyme is encoded by a gene mapped to 22q13 -,22qter. In order to determine the mCleC"lar nature of this enzyme, efforts were made to isolate the full-length cDNA and genomic sequences of the ellzyme. This enzyme was purified, oligonucleotideprobes were synthesized and a full-length cDNA was isolated. The 2156 bp cDNA encoded 411 amino acids. This enzyme was found to have nine exons and a single C .+ T transition was identified in the eighth won. The C--fT transition leaded to Argfhp at the 329th amino acid of the enzyme. This point mutation resulted in enzymaticallynon-active protein. Thus, this new disease ~8s elucidated to be caused by an exonic point mutation.
ULTRASTRUCTURALAND BIOCHEMICAL STUDIES OF A PATIENT WITH MENTAL RETARDATION AND MULTIPLE ANGIOMA. T.KANZAKIl, Y.KUNOl. Y.HIRASAYPSH12,AND N.G.FASER'. Deot. of Dermatol.. Nanova and Deot. of _- Citv - Univ.._Nanova .ii , -Bibch.rn Univ., of Shizuoka, Shizucka and Raigmore Hosp., scotw?. A Scot
female, 26 y/o, with mental retardation had multiple small angioma on her trunk which was recognized at 8 y/o (J.Med.Genet.). It was capillary hemangioma histologically. Electron microscopic studies disclosed many clear membrane-limitedcytoplasmic vacuoles in endathelial cells, pericytes and dermal fibroblasts. These clear vac"oles have been reported to be observed in various lysosomal storage diseases, such as mucopolysaccharidoses.glyccproteinosesor glycolipidoses. Enzymatic activities of lysosomal enzymes in her serum were within normal limits. Those were A-,Pgalactosidase,slalidase,j-L-fucosidase, d-N-acetylgalactosaminidase,d-rrannosidase,P-glucuronidase,6 -glucosidase and others. Biochemical snalybea with thin layer chromatographies showed no abnormality in glycopeptidesbut disclosed abnormal ganliosides exretion in her urine. These results suggest that this patient appears to have novel gangliosidosiswith mental retardation and multiple capillary hemangioma.
The possible role of fat cellsin the pathogenesis cl cutaneous leSiCnSOf Behcet'ssyndrome: Animmunoelectron microscopicstudy
DongsikBang,MD andSungnackLee.MD Departmentof Dermatology,Yonsei University Seoul. Korea
AUTOANTIBODY TO CARBONIC ANHYDRASE IN AUTOIMMUNE DISEASE SERA -ITS POSSIBLE CLINICAL AND PATHOGENICAL SIGNIFICANCEYASUNORI INAGAKI, YOKO YOSHIDA AND HIROAKI UEKI Department of Dermatology, Kawasaki Medical School, Kurashlki Carbonic anhydrase (CA) is an extremely basic and important enzyme for a regulation of acid-base status in viva with a wide phyletic distribution. Several different classes of this enzyme are known, and, at least, four enzymi’c forms were designated in humans. However, their amino acid sequences show a great deal of homology. CA are widely distributed in mamJKIli.3” tissues such as erythrocytes, gastric mucosa, renal cells, epidermal cells and so on. We found autoantibody reactive with CA, in sera from patients with several autoimmune disease, using the immunoblotting and the ELISA method. The incidence of the antibody “as not high among the patients. Thus we can conclude that occurrance of the autoantibody reactive with such a” important and fundamental enzyme is a significant phenomenon in autoimmune disorders.
PURIFICATION ANTIGEN YOSHINAO
Collegeof Medicine,
AND
NURO,
Department
attention was paidtothe appearance
of lymphocyte-macrophage-fat-cell adhesiOn and fat-cell lysisin the lobulesOf the subcutaneous fat of the erythema nodosum-like lesions of Behcet's syndrome. In this study, biopsy specimens from 10 patientswith typicalBehcet's syndrome were examined by immunoelectrcn microscopy with PaI'fiCUlar attentiontothechange of fat
cells
OF A HUNAN
DNA BINDING
NASARU
CENTRONERE
PROTEIN
OHASHI.
of Dermatology,
Nagoya
Univ.
School
of Medicine,
Nagoya Anticentromere Investlgatc
We have recentlyreported that special
CHARACTERIZATION
AS AN ALPHOID
et al.
reported
antigens, Hela
the
CENP-B,
satellite)(J. from
antibody
has provided
the biochemistry
Call
nuclear
interaction
and
of one
an alphoid
Biol.109:1963). extract
a probe
of the centramere. DNA We
to Nasumoto
of centromere
(a human
centromeric
purified
by its alpboid
DNA
CENP-Ei
binding
activity. Q-sepharose and alphoid DNA column were used, and fractions were assayed by DNA lmmunoprscipitation and immunoblottlng in each step of the purification. Specific activity of alphoid DNA blndinn was Increased over ten thousand fold. The resultsof DNaseI footprinting and south-western blot suggested that CENP-B itself has the activity to bind to a specific sequence of an alphoid DNA. We conclude that CENP-B is a specific DNA binding protein.